Formulations of bendamustine

ABSTRACT

Methods of treatment using bendamustine formulations designed for small volume intravenous administration are disclosed. The methods conveniently allow shorter administration time without the active ingredient coming out of solution as compared to presently available formulations.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority from U.S. ProvisionalPatent Application Ser. Nos. 61/613,173, filed Mar. 20, 2012, and61/669,889, filed Jul. 10, 2012, the disclosure of each of which isincorporated herein by reference.

BACKGROUND OF THE INVENTION

Bendamustine is used in the treatment of a number of cancers includingleukemias, Hodgkin's disease and multiple myelomas. Bendamustine(present as the HCl salt) is the active ingredient of the commercialproduct Treanda™, a lyophilized powder for reconstitution. Currentlabeling requirements call for the reconstituted product to beimmediately (within 30 minutes) diluted into 500 mL of parenterallyacceptable diluents such as 0.9% saline (normal saline) or 2.5%dextrose/0.45% saline and administered as part of an intravenousinfusion delivering 100 mg/m² over 30 minutes or 120 mg/m² over 60minutes. The diluted admixture may be stored at 2-8° C. for up to 24hours, or 3 hours at room temperature (15-30° C.); administration mustbe completed within this period due to limited chemical stability inaqueous solutions.

Solubility limitations at 2-8° C. with currently approved and/oravailable formulations are believed to prevent current formulations frombeing administered in smaller more concentrated infusion volumes up toabout 150 ml; at volumes below 150 ml, solubility is not sufficient evenat 25° C. Side effects associated with extravasation and local erythema,swelling and pain at the injection site also dictate that the infusionbe as dilute as possible. Therefore, precautions are taken to avoidextravasation, including monitoring of the intravenous infusion site forredness, swelling, pain, infection, and necrosis during and afteradministration of bendamustine. Higher infusion volume and longerinfusion times, however, are associated with many drawbacks. Forexample, after reconstitution, the current product has a short period ofstability, degradation of the drug occurs from the time ofreconstitution until the entire large volume infusion has beencompletely administered. The current label for Treanda™ thereforeinstructs that the admixture should be prepared as close as possible tothe time of patient administration, and that administration of Treanda™must be completed within the durations indicated above. From patientcomfort and nursing administration points of view, higher infusionvolumes and long infusion times are undesirable. Higher infusion volumesmay be associated with higher likelihood of weight gain and edema.Shorter infusion times and smaller infusion volumes result in a betterquality of life experience for the patient by reducing the overall“stress” to the patient and reducing the time spent in the infusionclinic. Shorter infusion times (and smaller volumes) also reduce thepotential extravasation (and shorten the patient monitoring timerequired). It would be advantageous if the drug could be administered insmaller volumes and over shorter times. The present invention addressesthese needs.

SUMMARY OF THE INVENTION

In a first aspect of the invention there are provided methods oftreating or preventing cancer or malignant disease in a subject such asa human. The methods include parenterally administering a volume ofabout 325 ml or less of a liquid composition containing:

a) from about 0.05 to about 12.5 mg/ml of bendamustine or apharmaceutically acceptable salt thereof;

b) a solubilizer comprising polyethylene glycol and propylene glycol;and optionally

c) a parenterally acceptable diluent;

over a substantially continuous period of less than or equal to about 30minutes to a subject in need thereof.

In alternative aspects of the invention there are provided methods oftreating or preventing a bendamustine-responsive condition in a subjectsuch as a human. In a first embodiment the methods include administeringless than or equal to 325 ml of a liquid composition which contains

Concentration Ingredient Range (mg/ml) Bendamustine HCl 0.05 to 1.6Solubilizer 1 propylene glycol 0.30 to 6.5 Solubilizer 2 PEG 400  3.3 to65 Monothioglycerol  0.02 to 0.35 NaOH  0.0 to 0.01over a substantially continuous period of less than or equal to about 30minutes to a subject in need thereof.

In a related second embodiment of this aspect of the invention, themethods include administering less than or equal to 325 ml of a liquidcomposition which contains

Concentration Ingredient Range (mg/ml) Bendamustine HCl 1.1 to 12.5Solubilizer 1 propylene glycol 4.5 to 51  Solubilizer 2 PEG 400 45 to500 Monothioglycerol 0.2 to 2.5  NaOH 0.0 to 0.04over a substantially continuous period of less than or equal to about 30minutes to a subject in need thereof.

The methods of the present invention take advantage of the fact that theconcentration of the bendamustine HCl is below the room temperaturesolubility limit of the vehicle into which it is placed. As a result,the bendamustine does not precipitate during administration to thepatient thereby substantially avoiding the side effects which wouldotherwise occur during small volume administration of therapeutic dosesof the drug. In addition, patients or subjects withbendamustine-responsive conditions can be treated using substantiallysmaller parenteral volumes which are well below the standard 500 mladministration volume.

DETAILED DESCRIPTION OF THE INVENTION

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art to which this invention belongs. In the event that there is aplurality of definitions for a term herein, those in this sectionprevail unless stated otherwise.

In a first aspect of the invention there are provided methods oftreating or preventing cancer or malignant disease in a subject orpatient who is preferably a human. The methods generally includeparenterally administering a volume of about 325 ml or less of a liquidcomposition containing:

a) from about 0.05 to about 12.5 mg/ml of bendamustine HCl or apharmaceutically acceptable salt thereof, the HCl salt being preferred;

b) a solubilizer comprising polyethylene glycol and propylene glycol;and optionally

c) a parenterally acceptable diluent;

over a substantially continuous period of less than or equal to about 30minutes to a subject in need thereof.

The solubilizer portion of the formulation preferably includes fromabout 0.3 to about 45% volume polyethylene glycol (PEG) and from about0.03 to about 5% volume propylene glycol (PG), as calculated on thebasis of the total or final volume administered. Stated alternatively,the final concentration of the PEG generally ranges from about 3 toabout 500 mg/ml, while the final concentration of the PG generallyranges from about 0.5 to about 51 mg/ml. Within these general ranges,certain aspects of the invention include concentration ranges for thePEG of from about 45 to about 500 mg/ml or from about 3.3 to about 63.3mg/ml; and for the PG ranges of from about 4.7 to about 50.6; or fromabout 0.02 to about 6.5 mg/ml.

In some aspects of the invention, the bendamustine is administeredintravenously as part of an intravenous infusion. Contemplated infusionvolumes are preferably less than 325 ml with volumes such as about 250ml, 100 ml, and 50 ml, with each volume varying about +/−10% or +1-15%being preferred in some embodiments. In alternative aspects of theinvention, the intravenous administration volume is suitable for IVbolus administration and may also include an amount of pharmaceuticallyacceptable diluent such as normal saline or one of the other diluentsdescribed herein which does not cause the solubility of the vehicle tofall below the concentration of the bendamustine. Stated alternatively,the final concentration of the bendamustine will be below the solubilityof the combination vehicle containing the mixture of propylene glycoland PEG and diluent. While most aspects of the invention are describedin the context of administering less than about 325 ml including allvehicle ingredients, excipients, etc., it should be appreciated thatvolumes as low as a few milliliters, e.g. about 2, can be used so longas the vehicle includes sufficient solubilizers to preserve thesolubility of the bendamustine therein during administration to thepatient.

For purposes of the present invention, the word “about” when used tomodify infusion volumes or concentrations shall be understood to includevalues which may vary by amounts of about +/−10% or 15%. In certainembodiments where the infusion volume is about 50 ml, the concentrationof the bendamustine HCl or other pharmaceutically acceptable saltthereof is preferably from about 0.5 to about 5.6 mg/ml. In embodimentswhere the infusion volume is about 100 ml, the concentration of thebendamustine HCl or other pharmaceutically acceptable salt thereof canbe preferably from about 0.1 to about 3.2 mg/ml. Similarly, in someaspects of the invention where the infusion volume is about 250 ml, theconcentration of the bendamustine HCl or other pharmaceuticallyacceptable salt thereof is from about 0.05 to about 1.4 mg/ml.

The solubilizer is preferably a mixture of polyethylene glycol,hereinafter “PEG” and propylene glycol, hereinafter “PG”. Thesolubilizer can also optionally include an antioxidant such asmonothioglycerol. The amount of antioxidant included is a formulationstabilizing amount, which, in the case of monothioglycerol ranges fromabout 2 to about 10 mg/ml. The PEG preferably has a molecular weight ofabout 400, i.e. PEG 400. Other molecular weight PEG's known to those ofordinary skill can be included if desired in alternative embodiments.

Certain aspects of the invention call for the ratio of the PEG to PGfound in the solubilizer to be about 90:10. In alternative aspects, theratio of the PEG to PG is about 85:15.

In some aspects of the invention, the total amount of solubilizer, i.e.blend of PEG and PG, included in infusion volumes of about 100-115 ml isfrom about 0.5 to about 26.5% vol.; while amounts of from about 0.2 toabout 5% vol. for the solubilizer are preferably included in infusionvolumes of about 250-265 ml; with solubilizer amounts of from about 2.0to about 22.4% vol. included in infusion volumes of about 50-65 ml.

Since the solubilizer is a blend, the amount of PEG and PG in variousvolumes (calculated as % vol.) can be as follows:

Solubilizer 50 ml 100 ml 250 ml PEG 20.12 11.33 4.9 PG 2.24 1.26 0.54

In some preferred embodiments, the methods of the invention areadvantageously carried out using bendamustine HCl containingcompositions administered as small volume infusions with volumes ofabout 50 ml or about 100 ml or about 250 ml. Such smaller volumes allowthe drug to be administered over a time period of about 10 minutes orless as part of an intravenous infusions containing a volume of about 50ml; about 15 minutes or less as part of an intravenous infusionscontaining a volume of about 100 ml or when volumes of about 250 ml areinfused, the IV infusion is administered over a time period of about 30minutes or less. Depending upon the amount of drug administered, the IVbolus volumes containing sufficient amount of the drug will be less than50 ml, with amounts of about 10 or 15 to 30 ml being sufficient.

The infusible compositions in many aspects of the invention will alsopreferably include the parenterally acceptable diluents such as waterfor injection (WFI), 0.9% saline (normal saline, preferred), 0.45%saline (half normal saline) or 2.5% dextrose/0.45% saline. Formulationswell suited for carrying out the methods described herein are alsodescribed in commonly assigned U.S. patent application Ser. No.13/016,473, filed Jan. 28, 2011, and Ser. No. 13/767,672 filed Feb. 14,2013, the contents of which are incorporated herein by reference. Asreviewed in the '672 patent application, some preferred bendamustineformulations can also include a minor amount of a pH adjuster such assodium formate, sodium phosphate, potassium hydroxide, phosphoric acidor, preferably, sodium hydroxide.

In an alternative embodiment of the invention, the bendamustineformulations used in the methods described herein can be one or more ofthose described in U.S. Pat. Nos. 8,344,006 and 8,076,366; and US PatentApplication Nos. 2013/0041004; 2012/0071532; 2010/0216858; 2006/0159713;and 2013/0041003, the contents of each of which are incorporated hereinby reference. It being understood that the vehicle into which thebendamustine HCl is placed will have sufficient bendamustine solubilitywhich exceeds the concentration of the drug included therein.

If desired, a sufficient amount of a concentrated, ready to use liquidformulation such one containing 25 mg/ml bendamustine HCl and alreadyadmixed with sufficient solubilizers can be transferred to a suitablefixed volume diluent container such as a bag containing 50, 100, 250 mlnormal saline or the like. Alternatively, lyophilized bendamustine HClcan be reconstituted, combined with sufficient solubilizer blends asdescribed herein and administered in accordance with the inventivemethods. In such embodiments, the actual amount delivered to the patientwill be slightly more than the diluent amount so as to allow for theaddition of the drug/solubilizer vehicle.

In some aspects of the invention, there are provided methods of treatingor preventing chronic lymphocytic leukemia (CLL). The small volumeinfusions can be given as part of any treatment protocol for whichbendamustine is included. Thus, the compositions described herein can beadministered as part of a poly-pharmaceutical treatment regimenaccording to known protocols with the exception that the concentratedbendamustine compositions described herein are administered in smallerinfusion volumes over significantly shorter administration periods. Forexample, some CLL treatment regimens can include administering thecompositions described herein intravenously as part of about 100 mlinfusions in about 20 minutes or less and more preferably in about 15minutes or less on days 1 and 2 of a 28 day cycle and repeating thecycle up to 6 times or longer if clinically appropriate. If 250 mlvolumes are used to deliver the bendamustine, the time of administrationis preferably about 30 minutes or less. If 50 ml volumes are used todeliver the bendamustine, the time of administration is preferably about10 minutes or less.

In spite of the smaller volumes, the amount of bendamustine HCladministered to the patient in need thereof per dose (infusion orotherwise) in some preferred embodiments is about 100 mg/m². In somealternative aspects of the invention, the amount of bendamustine HCladministered to the patient in need thereof as part of the 50, 100 or250 ml infusion is an amount sufficient to provide a dosage of 50 or 25mg/m². Additional administration dosages will be apparent to those ofordinary skill based upon clinical experience, patient need withoutundue experimentation.

In other aspects of the invention, there are methods of treating orpreventing the malignant disease of indolent B-cell non-Hodgkin'slymphoma. In these aspects, the composition is administeredintravenously as a 100 ml infusion in less than 20 minutes and morepreferably in about 15 minutes or less on days 1 and 2 of a 21 day cyclefor up to 8 cycles or longer if clinically appropriate. If 250 mlvolumes are used to deliver the bendamustine, the time of administrationis preferably about 30 minutes or less. If 50 ml volumes are used todeliver the bendamustine, the time of administration is preferably about10 minutes or less. The amount of bendamustine administered to thesubject is preferably about 120 mg/m², although in alternativeembodiments, the amount administered ranges from about 90 or 60 mg/m².As will be appreciated, further alternative dosage amounts will beapparent to those of ordinary skill based upon clinical experience,patient need without undue experimentation.

It will be appreciated by those skilled in the art that theabove-mentioned dosages calculated in mg/m² for purposes of body surfacearea (BSA) are consistent with the bendamustine HCl concentrations alsodescribed herein, e.g. 0.5 to 5.6 mg/ml. In the alternative, theinvention also contemplates IV bolus administration ofbendamustine-containing formulations in volumes which can beadministered via syringe, e.g. from a few milliliters up to about 50milliliters, with therapeutic amounts of the drug in a concentrationwhich does not exceed the vehicle solubility for the drug therein.

Further embodiments of the invention include methods of treating orpreventing a bendamustine-responsive condition in a subject such as ahuman. In a first embodiment, the methods include administering lessthan or equal to 325 ml of a liquid composition which contains

Concentration Ingredient Range (mg/ml) Bendamustine HCl 0.05 to 1.6Solubilizer 1 propylene glycol 0.30 to 6.5 Solubilizer 2 PEG 400  3.3 to65 Monothioglycerol  0.02 to 0.35 NaOH  0.0 to 0.01over a substantially continuous period of less than or equal to about 30minutes to a subject in need thereof. More preferably, theadministration time is well below 30 minutes and the administration timewill decrease as the volume administered decreases.

Bendamustine formulations containing the above ingredients are capableof delivering approximately 25 mg of the drug as the HCl salt in volumesof pharmaceutically acceptable diluent ranging from about 325 ml down toabout 15 ml. For example, 1 ml of a bendamustine HCl ready to use liquidavailable from Eagle Pharmaceuticals containing

Concentration Ingredient (mg/ml) Bendamustine HCl 25 PG 103.2 PEG 4001013.4 Monothioglycerol 5 NaOH 0.08is combined with 300 ml of a normal saline diluent to provide a final IVinfusion containing 301 ml and a bendamustine final concentration of0.08 mg/ml.

One ml of the 25 mg/ml Eagle bendamustine HCl is diluted into additionaldiluent volumes as shown below:

Diluent Volume Final Volume Final Bendamustine (ml) (ml) Conc. (mg/ml)200 201 0.12 100 101 0.25 50 51 0.49 30 31 0.81 15 16 1.56

The measured solubility of the bendamustine HCl in thediluent/solubilizer combination (50 ml diluent plus 1 ml of 25 mg/mlbendamustine HCl and solubilizers, etc.) at room temperature was 10.5mg/ml using normal saline and 14.2 mg/ml using half normalsaline/dextrose. The solubility of the diluent/solubilizer combinationfar exceeded the bendamustine concentration, thus assuring the avoidanceof precipitated drug prior to or during administration. As will beappreciated by those of ordinary skill, as the concentration ofsolubilizers increases with respect to the total volume in smalladministration doses, the solubility of the bendamustine is maintained.

In a related second embodiment of this aspect of the invention, themethods include administering less than or equal to 325 ml of a liquidcomposition which contains

Concentration Ingredient Range (mg/ml) Bendamustine HCl 1.1 to 12.5Solubilizer 1 propylene glycol 4.5 to 51  Solubilizer 2 PEG 400 45 to500 Monothioglycerol 0.2 to 2.5  NaOH 0.0 to 0.04over a substantially continuous period of less than or equal to about 30minutes to a subject in need thereof. As was the case above, theadministration time will decrease with the decrease in volumeadministered.

Bendamustine formulations containing the above ingredients are capableof delivering approximately 360 mg of the drug as the HCl salt involumes of pharmaceutically acceptable diluent ranging from about 325 mldown to about 15 ml. As was the case above, the measured solubility ofthe bendamustine HCl in the diluent/solubilizer combination (1 mldrug+solubilizers, etc. and 50 ml diluent) at room temperature was 10.5mg/ml using normal saline and 14.2 mg/ml using half normalsaline/dextrose.

Instead of using only 1 ml of the above described Eagle 25 mg/mlbendamustine HCl ready to use liquid, 14.4 ml is combined with variousamounts of diluent.

Diluent Volume Final Volume Final Bendamustine (ml) (ml) Conc. (mg/ml)300 314.4 1.15 200 214.4 1.68 100 114.4 3.15 50 64.4 5.59 30 44.4 8.1115 29.4 12.24

In each case, the solubility of the diluent/solubilizer combinationexceeds the bendamustine concentration, thus assuring the avoidance ofprecipitated drug prior to or during administration.

EXAMPLES

The following examples serve to provide further appreciation of theinvention but are not meant in any way to restrict the effective scopeof the invention.

Example 1

The solubility of bendamustine HCl, obtained from two different sources,in 0.9% saline and 0.9% saline containing from different amounts of anon-aqueous solubilizer comprising a mixture of polyethylene glycol 400and propylene glycol (in the volume proportion of 90:10) with andwithout 5 mg/ml monothioglycerol was determined at both room temperature(22-23° C.) and at refrigerated temperature (5° C.). Essentially, anexcess of bendamustine HCl was added to solvents comprising of variousvolume percent of the non-aqueous solubilizer in 0.9% saline, andallowed to equilibrate with shaking for 30 minutes at room temperature,or for 24 hours at refrigerated temperature. At the end of theequilibration step, the suspensions were filtered through a 0.2 micronfilter to remove undissolved bendamustine, and the filtrate solutionsanalyzed for bendamustine HCl content using a HPLC assay; quantificationwas performed against a bendamustine HCl reference standard. Thesolubility data are presented in Table 1.

TABLE 1 Solubility of bendamustine HCl in 0.9% saline with variousamounts of non-aqueous solubilizer (90:10 PEG400:PG with and without 5mg/mL monothioglycerol (MTG)) Solubility of bendamustine HCl VolumeVolume % (mg/mL) in 90:10 PEG 400/PG % of of Non- Room temperature 5° C.Dilution Normal aqueous API API API Fold Saline Solubilizer Source ASource B* B Source A n/a 100.0 0.0 3.461 3.304 1.175 40 97.5 2.5 3.9873.889 nd 20 95.0 5.0 4.429 4.204 2.022 13.3 92.5 7.5 nd 4.742 nd 10 90.010.0 5.626 5.351 2.431 8 87.5 12.5 nd 5.825 nd 6.7 85.0 15.0 7.012 6.5542.900 5.7 82.5 17.5 nd 7.641 3.328 5 80.0 20.0 8.642 8.492 3.824 3.370.0 30.0 12.006  11.407 nd *solvent also contained 5 mg/mlmonothioglycerol nd = not determined; API = active pharmaceuticalingredient

Example 2

Bendamustine-containing compositions are prepared by adding 5 mg/ml ofthioglycerol to a mixture containing 90% polyethylene glycol 400 and 10%propylene glycol. As indicated in the Table 2 below, NaOH may be addedto the PEG in an amount sufficient to get apparent pH of greater than orequal to 6.5 as measured using the pH method outlined in the USPmonograph for polyethylene glycol (PEG). Bendamustine (BDM) is thenadded to the sample to a concentration of 10 mg/ml.

TABLE 2 Formulation BDM - 10 mg/mL Thioglycerol - 5 mg/mL PEG 400:PG(90:10) qs to 1 mL BDM - 10 mg/mL Thioglycerol - 5 mg/mL PEG 400:PG(90:10) qs to 1 mL (PEG 400 Treated with NaOH)

The compositions are then admixed with normal saline based on the totaldose of bendamustine HCl, which in turn is based on the patient bodysurface area (BSA) and the dosing regimen (100 mg/m² for CLL and 120mg/m² for NLL; although dose modifications of 90, 60, 50, and 25 mg/m²are possible, only the highest two dosing regimens are considered forillustrative purposes, as these result in the highest concentration ofbendamustine during infusion). The 100 ml infusion is then made byadmixing the dose appropriate volume of the 10 mg/ml solution with a 100ml portion of normal saline to provide an infusible compositioncontaining the appropriate dose of bendamustine (as the HCl salt) in thefinal admixture, which can be administered intravenously over about 15minutes to a patient in need thereof.

As seen in Table 3, the concentrations of bendamustine (as HCl salt) andthe corresponding volume percent of non-aqueous component are well belowthe corresponding solubilities at both room temperature and refrigeratedtemperature as detailed in Table 1. For example, for a 2.0 m² (average)patient dosed at 120 mg/m2, the final concentration of bendamustine HClin a 100 ml admixture is 1.94 mg/mL. This is above the solubility ofbendamustine HCl at refrigerated storage conditions in the absence ofany non-aqueous components (1.175 mg/ml as shown in Table 1 for 100%normal saline), as would be the case with the currently approvedTreanda™ product, thereby precluding preparation and storage of a 100 mladmixture volume at refrigerated conditions. However, the use of thenon-aqueous bendamustine formulation described in this example resultsin the presence of 19.4% of the non-aqueous component in the finaladmixture, which improves the solubility to about 3.8 mg/mL (solubilityof 3.824 mg/mL at 2-8° C. with 20% non-aqueous component, as shown inTable 1). Therefore, the solubility with the non-aqueous formulation iswell above the final concentration (of bendamustine HCl) of 1.94 mg/mL,allowing preparation and storage of the 100 ml admixture at refrigeratedconditions. In this example, the room temperature solubilities in 100%normal saline and 80% normal saline (with 20% non-aqueous component) areabout 3.3 mg/ml and 8.5 mg/ml, respectively (see Table 1), which arealso well above the final concentration of 1.94 mg/ml. Therefore, 100 mladmixtures of the non-aqueous formulation described in the example mayalso be prepared and stored at room temperature. In addition, thenon-aqueous formulation of bendamustine described in this example atTable 2 may be diluted into smaller infusion volumes ranging from 250 mlor less, and stored at either room temperature or refrigeratedtemperature, with bendamustine continuing to remain in solution forextended periods of time as compared to currently availableformulations.

TABLE 3 Concentrations of bendamustine (BDM, as HCl salt) andcorresponding volume % of non-aqueous (NA) component in the finaladmixture, for volumes ranging from 100 ml to 250 ml For 10 mg/mLFormulation 100 mg/m² 120 mg/m² 100 mg/m² 120 mg/m² dose dose dose doseBDM BDM BDM BDM BSA Conc % NA Conc % NA Conc % NA Conc % NA (m²) (mg/ml)comp. (mg/ml) comp. (mg/ml) comp. (mg/ml) comp. Admix. Volume 250 mLAdmix. Volume 200 mL 1.00 0.38 3.8 0.46 4.6 0.48 4.8 0.57 5.7 1.25 0.484.8 0.57 5.7 0.59 5.9 0.70 7.0 1.50 0.57 5.7 0.67 6.7 0.70 7.0 0.83 8.31.75 0.65 6.5 0.77 7.7 0.80 8.0 0.95 9.5 2.00 0.74 7.4 0.88 8.8 0.91 9.11.07 10.7 2.25 0.83 8.3 0.97 9.7 1.01 10.1 1.19 11.9 2.50 0.91 9.1 1.0710.7 1.11 11.1 1.30 13.0 2.75 0.99 9.9 1.17 11.7 1.21 12.1 1.42 14.23.00 1.07 10.7 1.26 12.6 1.30 13.0 1.53 15.3 Admix. Volume 150 mL Admix.Volume 100 mL 1.00 0.63 6.3 0.74 7.4 0.91 9.1 1.07 10.7 1.25 0.77 7.70.91 9.1 1.11 11.1 1.30 13.0 1.50 0.91 9.1 1.07 10.7 1.30 13.0 1.53 15.31.75 1.04 10.4 1.23 12.3 1.49 14.9 1.74 17.4 2.00 1.18 11.8 1.38 13.81.67 16.7 1.94 19.4 2.25 1.30 13.0 1.53 15.3 1.84 18.4 2.13 21.3 2.501.43 14.3 1.67 16.7 2.00 20.0 2.31 23.1 2.75 1.55 15.5 1.80 18.0 2.1621.6 2.48 24.8 3.00 1.67 16.7 1.94 19.4 2.31 23.1 2.65 26.5

Example 3

The procedures of Example 2 are repeated except that the doseappropriate volume of the 10 mg/ml bendamustine solution is diluted into250 ml of normal saline. The final concentration of bendamustine in the250 ml volume container ranges from about 0.05 mg/ml to about 1.3 mg/ml.

Example 4

The approximately 100 ml bendamustine HCl infusion of Example 2 isadministered to a patient in about 15 minutes.

Example 5

Bendamustine-containing compositions may be prepared by adding 5 mg/mlof thioglycerol to 90% polyethylene glycol 400 and 10% propylene glycol.As indicated in the Table 4 below, NaOH may be added in an amountsufficient to get apparent pH of greater than or equal to 6.5 asmeasured using the pH method outlined in the USP monograph forpolyethylene glycol (PEG). Bendamustine is then added to the sample to aconcentration of 25 mg/ml as indicated in Table 4 below.

TABLE 4 Formulation BDM - 25 mg/mL Thioglycerol - 5 mg/mL PEG 400:PG(90:10) qs to 1 mL BDM - 25 mg/mL Thioglycerol - 5 mg/mL PEG 400:PG(90:10) qs to 1 mL (PEG 400 Treated with NaOH)

The compositions are then admixed with normal saline based on the totaldose of bendamustine HCl, which in turn is based on the patient bodysurface area (BSA) and the dosing regimen (100 mg/m² for CLL and 120mg/m2 for NLL; although dose modifications of 90, 60, 50, and 25 mg/m²are possible, only the highest two dosing regimens are considered forillustrative purposes, as these result in the highest concentration ofbendamustine during infusion). Table 5 below provides the finalconcentration of bendamustine (as the HCl salt) in the final admixture,for volumes ranging from 250 ml to 50 ml.

As seen in Table 5, the concentrations of bendamustine (as HCl salt) andthe corresponding volume percent of non-aqueous component are well belowthe corresponding solubilities at room temperature as detailed in Table1, for all admixture volumes up to 50 ml. For example, for a 2.0 m²(average) patient dosed at 120 mg/m², the final concentration ofbendamustine HCl in a 50 ml admixture is 4.03 mg/ml. This is above thesolubility of bendamustine HCl at both refrigerated and room temperatureconditions in the absence of any non-aqueous components (1.175 mg/ml at2-8° C. and 3.304-3.461 mg/ml at room temperature, as shown in Table 1for 100% normal saline), as would be the case with the currentlyapproved Treanda product, thereby precluding preparation and storage ofa 50 ml admixture volume. However, the use of the non-aqueousbendamustine formulation described in this example results in thepresence of 16.1% of the non-aqueous component in the final admixture,which improves the room temperature solubility to about 6.5 mg/ml(solubility of 6.554 mg/ml and 7.012 mg/ml with 15% non-aqueouscomponent, as shown in Table 1). Therefore, the solubility with thenon-aqueous formulation is well above the final concentration (ofbendamustine HCl) of 4.03 mg/mL, allowing preparation and storage of the100 ml admixture at room temperature conditions. Therefore, thenon-aqueous formulation of bendamustine described in this example may bediluted into smaller infusion volumes ranging from 250 ml or less, withbendamustine continuing to remain in solution if maintained at roomtemperature. However, at refrigerated temperatures, the concentrationsof bendamustine (as HCl salt) and the corresponding volume percent ofnon-aqueous component exceed the corresponding solubilities as detailedin Table 1, for all admixture volumes equal to or below 150 ml. In thescenario above, the solubility at refrigerated conditions with 15%non-aqueous component has improved to 2.9 mg/ml but is still below thefinal concentration of 4.03 mg/ml. Therefore, 50 ml admixtures of thenon-aqueous formulation described in the example cannot be prepared andstored at refrigerated temperatures. However, for a 150 ml admixture,the final concentration of bendamustine HCl in this scenario is 1.5mg/ml with about 6.0% non-aqueous component, which is below thesolubility limit (of 2.022 mg/ml at 5% non-aqueous at 2-8° C.).Therefore, the non-aqueous formulation of bendamustine described in thisexample may be diluted into smaller infusion volumes ranging from 250 mlto 50 ml, and stored at only room temperature (but not refrigeratedtemperature), with bendamustine continuing to remain in solution. Forstorage at refrigerated temperatures, the minimum admixture volume thatcan be used is 150 ml or higher.

TABLE 5 Concentrations of bendamustine (BDM, as HCl salt) andcorresponding volume % of non-aqueous (NA) component in the finaladmixture, for volumes ranging from 100 ml to 250 ml For 25 mg/mLFormulation 100 mg/m² 120 mg/m² 100 mg/m² 120 mg/m² dose dose dose doseBDM BDM BDM BDM BSA Conc % NA Conc % NA Conc % NA Conc % NA (m²) (mg/ml)comp. (mg/ml) comp. (mg/ml) comp. (mg/ml) comp. Admix. Volume 250 mLAdmix. Volume 200 mL 1.00 0.39 1.6 0.47 1.9 0.49 2.0 0.59 2.3 1.25 0.492.0 0.59 2.3 0.61 2.4 0.73 2.9 1.50 0.59 2.3 0.70 2.8 0.73 2.9 0.87 3.51.75 0.68 2.7 0.81 3.3 0.85 3.4 1.01 4.0 2.00 0.78 3.1 0.92 3.7 0.96 3.81.15 4.6 2.25 0.87 3.5 1.04 4.1 1.08 4.3 1.28 5.1 2.50 0.96 3.8 1.15 4.61.19 4.8 1.42 5.7 2.75 1.05 4.2 1.25 5.0 1.30 5.2 1.55 6.2 3.00 1.15 4.61.36 5.4 1.42 5.7 1.68 6.7 Admix. Volume 150 mL Admix. Volume 100 mL1.00 0.65 2.6 0.78 3.1 0.96 3.8 1.15 4.6 1.25 0.81 3.2 0.96 3.8 1.19 4.81.42 5.7 1.50 0.96 3.8 1.15 4.6 1.42 5.7 1.68 6.7 1.75 1.11 4.5 1.33 5.31.64 6.5 1.94 7.7 2.00 1.27 5.1 1.50 6.0 1.85 7.4 2.19 8.8 2.25 1.42 5.71.68 6.7 2.06 8.3 2.44 9.7 2.50 1.56 6.3 1.85 7.4 2.27 9.1 2.68 10.72.75 1.71 6.8 2.02 8.1 2.48 9.9 2.92 11.7 3.00 1.85 7.4 2.19 8.8 2.6810.7 3.15 12.6 Admix. Volume 50 mL 100 mg/m² dose 120 mg/m² dose BSA BDMConc % NA BDM Conc % NA (m²) (mg/ml) comp. (mg/ml) comp. 1.00 1.85 7.42.19 8.8 1.25 2.27 9.1 2.68 10.7 1.50 2.68 10.7 3.15 12.6 1.75 3.07 12.33.60 14.4 2.00 3.45 13.8 4.03 16.1 2.25 3.81 15.3 4.44 17.8 2.50 4.1716.7 4.84 19.4 2.75 4.51 18.0 5.22 20.9 3.00 4.84 19.4 5.59 22.4

Example 6

The hemolytic potential of the non-aqueous bendamustine formulationindicated in Table 4 (Example 5), when admixed with 250 ml and 100 ml ofnormal saline, was assessed. The hemolysis study was conducted at thehighest final bendamustine HCl concentrations expected at theseadmixture volumes, namely, for a 3.0 m² patient dosed at 120 mg/m². Atthis dosing, the final bendamustine HCl concentration for 250 ml and 100ml admixture volumes is 1.36 mg/ml and 3.15 mg/ml, respectively (Table5). Human whole blood (1 ml) was incubated at 37° C. for approximately30 minutes with admixed bendamustine HCl solutions at 1.4 mg/ml or 3.2mg/ml at blood to drug solution volumetric ratios of 1:2 and 1:1,respectively. These volumetric ratios correspond to infusion times of 15minutes and 10 minutes, respectively, for the 250 ml and 100 mladmixture volumes. A placebo of the bendamustine formulation (withoutthe active ingredient) was also evaluated at these concentrations andvolumetric ratios. A positive control (1% saponin solution), a negativecontrol (normal saline), and Treanda™ diluted in normal saline to thehighest concentration stated in the prescribing information (0.6 mg/ml)were included in the study. Following incubation and centrifugation ofthe samples, the plasma was harvested and hemolysis was evaluated byspectrophotometric analysis for hemoglobin in the supernatant. Theresults are summarized in Table 6. No hemolysis was observed with thenon-aqueous bendamustine formulation when diluted with saline at eitherconcentration or volumetric (blood:drug solution) ratios, or with thecorresponding placebo at comparable sample volumes; supernatants fromall samples were light yellow. In conclusion, no hemolytic effects areobserved with non-aqueous bendamustine formulations when diluted tosmaller volumes (100 to 250 ml) and infused in shorter times (10-15minutes) than current practice.

TABLE 6 Hemolytic Potential test results Mixture Human Hemoglobin^(a)Supernatant Tube blood plus: (mg/dL) Test Result Color^(b) No. TestArticle A (25 mg/mL, diluted to 3.2 mg/mL with saline) - 100 mladmixture 1 N Light yellow 1 0 N Light yellow 2 1 N Light yellow 3 TestArticle A (25 mg/mL, diluted to 1.4 mg/mL with saline) - 250 mladmixture 2 N Light yellow 4 1 N Light yellow 5 1 N Light yellow 6 TestArticle A Vehicle (Placebo, diluted with 100 ml saline) 0 N Light yellow7 1 N Light yellow 8 0 N Light yellow 9 Test Article A Vehicle (Placebo,diluted with 250 ml saline) 1 N Light yellow 10 4 N Light yellow 11 1 NLight yellow 12 Treanda ™ (5 mg/mL, diluted to 0.6 mg/mL with saline) 3N Light yellow 43 4 N Light yellow 44 2 N Light yellow 45 NegativeControl (normal saline) 9 N Yellow 55 5 N Yellow 56 3 N Yellow 57Positive Control (1% Saponin)^(c) 5949 P Red 58 5974 P Red 59 6386 P Red60 N = Negative, no hemolysis. NA = Not applicable. P = Positive,hemolysis. ^(a)Hemoglobin index of the mixture supernatants. ^(b)Plasmaseparated from whole blood plasma. ^(c)1% Saponin. Saponin is ahemolytic agent used to lyse erythrocytes.

Example 7

The local tolerance (intravenous (IV) and perivascular (PV)) of thenon-aqueous bendamustine-containing composition indicated in Table 4(Example 5), when admixed with 100 ml of normal saline and infused over10 minutes, was assessed. New Zealand White rabbits (3 males [IV] and 2males [PV]) received a single dose of bendamustine formulation (admixedwith 100 ml saline to a final concentration of 3.2 mg/ml bendamustineHCl) and corresponding placebo in the left and right ear, respectively.The formulation was administered as either intravenous infusion (5 mg/kgin 10 minutes), or perivascular injection (250 μA) to determine localtolerance. Treanda™ reconstituted and admixed with normal saline to afinal concentration of 0.6 mg/ml (the highest concentration stated inthe label) was also studied either as a 30 minute IV infusion (theshortest infusion time stated in the label), as well as perivascularinjection (250 μA). Animals were held for a 96 hour (post-dose)observation period. During the observation period, dermal scores wererecorded for all administration sites. At the end of the observationperiods, animals were euthanized and a macroscopic and microscopicexamination of both ears was performed. Parameters evaluated during thestudy were: viability, clinical observations, body weights, macroscopicobservations and microscopic pathology.

The results of the local tolerance study are summarized in Table 7 (inlife dermal observations) and Table 8 (microscopic pathology forperivascular administration).

In Life Dermal Observations:

As seen in Table 7, there was transient, dermal irritation in the formof slight to moderate erythema and moderate edema noted between 24 and72 hours post dose, in each of the groups receiving eitherbendamustine-containing formulations or placebo material intravenously.At 96 hours, irritation was limited to a few individual sites treatedwith test or placebo articles. Only a limited number of animals wereaffected, and there was no consistent pattern of irritation within adose group (either for test article or placebo). The bendamustineformulations were considered not to produce dermal irritation whenadministered intravenously.

Perivascular administration of bendamustine formulations (0.25 mlinjection volume) produced dermal irritation in all groups. Local signsof dermal irritation following perivascular administration were mostlycharacterized by slight (group 6—Treanda™) or slight to moderate (group7—non aqueous bendamustine formulation of example 5) erythema, andslight edema (groups 7). The severity of the irritation observedcorrelated with the dose and/or concentration of the test articleadministered, with placebo groups generally showing a lesser level ofirritation than the corresponding test-article formulation.

TABLE 7 Summary of in life dermal observations Dermal observationsduring 96 hour post dose period (incidence and most severe level oferythema and edema noted) Left ear Right ear (Bendamustine-containing(Placebo Material - Left ear/Right ear formulation) material)Intravenous administration Group 1: Treanda ™ diluted — (2/3) to 0.6mg/ml in saline/ Slight Treanda Placebo - 500 ml admixture Group 2:Non-aqueous — (1/3) bendamustine formulation Moderate 25 mg/ml dilutedto 3.2 mg/ml in saline/placebo (+saline) - 100 ml admixture Perivascularadministration Group 6: Treanda ™ diluted (1/2) (1/2) to 0.6 mg/ml insaline/ Slight Slight Treanda ™ Placebo - 500 ml admixture Group 7:Non-aqueous (2/2) (2/2) bendamustine formulation Moderate Slight 25mg/ml diluted to 3.2 mg/ml in saline/placebo (+saline) - 100 mladmixture

Microscopic Pathology:

Intravenous administration of test articles/placebos was generally welltolerated; no test article related effects were observed. Perivascularadministration of bendamustine-containing formulations (includingTreanda™) was associated with dose and/or concentration related minimalto marked edema/collagen degeneration and mixed inflammation inperivascular tissues. The non-aqueous formulation of bendamustine (Group7) was nominally more severe in grade than Treanda™ (Group 6).

TABLE 8 Incidence and Average Severity of Microscopic Findings atPerivascular Sites Formulation Treanda ™ Non Aqueous (diluted toBendamustine 0.6 mg/ml)/ (diluted to Treanda ™ 3.2 mg/ml)/ PlaceboPlacebo Group number 6 7 No. Animals examined 2 2 RIGHT EAR (RE) -Placebo Incidence (Average Severity)* RE Injection site Hemorrhage 0(0.0) 0 (0.0) Mixed Inflammation 0 (0.0) 0 (0.0) Edema/CollagenDegeneration 1 (0.5) 0 (0.0) Degeneration/Inflammation, 0 (0.0) 0 (0.0)Vascular RE 2 cm distal Edema/Collagen Degeneration 0 (0.0) 0 (0.0)Degeneration/Inflammation, 0 (0.0) 0 (0.0) Vascular RE 4 cm distalDegeneration/Inflammation, 0 (0.0) 0 (0.0) Vascular LEFT EAR (LE) - TestArticle Incidence (Average Severity) LE Injection site Hemorrhage 0(0.0) 0 (0.0) Mixed inflammation 0 (0.0) 2 (2.0) Edema/CollagenDegeneration 1 (1.0) 2 (1.5) Epidermis, Crust/Pustule, 0 (0.0) 0 (0.0)Erosion/Ulceration Degeneration/Inflammation, 1 (1.5) 1 (1.5) VascularLE 2 cm distal Hemorrhage 0 (0.0) 0 (0.0) Mixed Inflammation 1 (0.5) 1(1.0) Edema/Collagen Degeneration 0 (0.0) 2 (2.0) Epidermis,Crust/Pustule, 0 (0.0) 0 (0.0) Erosion/UlcerationDegeneration/Inflammation, 0 (0.0) 0 (0.0) Vascular LE 4 cm distalHemorrhage 0 (0.0) 0 (0.0) Mixed inflammation 0 (0.0) 1 (1.5)Edema/Collagen Degeneration 0 (0.0) 1 (2.0) Epidermis, Crust/Pustule, 0(0.0) 0 (0.0) Erosion/Ulceration Degeneration/Inflammation, 0 (0.0) 0(0.0) Vascular *The number in parentheses represents the averageseverity score; the total of severity scores of the findings divided bythe number of animals in the group.

Conclusion:

No test-article related irritation effects were observed for thenon-aqueous formulation of bendamustine via the IV route, indicatingthat proper administration of this formulation did not result in anyadverse local reaction. Perivascular administration of the non-aqueousbendamustine formulation, which is primarily related to effects that mayoccur if extravasation should occur, resulted in irritation that wasgenerally comparable to Treanda. Therefore, the non-aqueous formulationof bendamustine described herein is well tolerated, despite the higherconcentration of the smaller infusion volume preparation.

Example 8

The chemical stability of the non-aqueous bendamustine formulation (25mg/ml) indicated in Table 4 (Example 5), when admixed with 50 ml and 100ml of normal saline, was assessed. For each admixture volume, theadmixture solutions were prepared at the expected lowest concentration(corresponding to a 1.0 m² patient dosed at 25 mg/m²) and the highestconcentration (corresponding to a 3.0 m² patient dosed at 120 mg/m²) ofbendamustine HCl in the final admixture. For the 50 ml admixture volume,the tested minimum and maximum concentrations are about 0.5 mg/ml and6.0 mg/ml, respectively. For the 100 ml admixture volume, the testedminimum and maximum concentrations are about 0.25 mg/ml and 3.2 mg/ml,respectively. The chemical stability of Treanda™ was also determined atthe lowest (0.2 mg/ml) and the highest (0.6 mg/ml) admixedconcentrations stated in the label. The chemical stability was monitoredat room temperature at periodic intervals up to 24 hours using avalidated HPLC assay. The results are summarized in Table 9.

TABLE 9 Dilution (Admixed) Stability of Bendamustine Formulations inNormal Saline at Room Temperature Attribute Highest Concentration (3.2mg/ml) Lowest Concentration (0.25 mg/ml) Time Initial 1 hr 3 hrs 6 hrs24 hrs Initial 1 hr 3 hrs 6 hrs 24 hrs Formulation Non AqueousBendamustine Formulation 25 mg/ml admixed with 100 ml normal salineAssay (mg/ml) 3.155 3.090 3.060 3.085 2.895 0.240 0.234 0.229 0.2240.196 Assay (% Initial) 100.0 97.9 97.0 97.8 91.8 100.0 97.5 95.4 93.381.7 Impurity - MCE (%) BLQ BLQ BLQ BLQ BLQ ND ND ND ND ND Impurity -HP1 (%) 0.244 0.606 1.237 2.236 6.707 0.525 1.449 3.495 5.529 13.424Impurity - Dimer (%) BLQ BLQ BLQ 0.068 0.158 ND ND ND BLQ 0.063 Singleunknown (%) 0.086 0.061 BLQ BLQ 0.098 ND ND ND ND BLQ Total (%) 0.330.67 1.24 2.30 6.96 0.58 1.45 3.50 5.53 13.49 Attribute HighestConcentration (6.4 mg/ml) Lowest Concentration (0.5 mg/ml) Time Initial1 hr hrs 6 hrs 24 hrs Initial 1 hr 3 hrs 6 hrs 24 hrs Formulation NonAqueous Bendamustine Formulation 25 mg/ml admixed with 50 ml normalsaline Assay (mg/ml) 6.62 6.60 6.60 6.54 6.46 0.475 0.470 0.455 0.4450.394 Assay (% Initial) 100.0 99.7 99.7 98.8 97.8 100.0 98.9 95.8 93.782.9 Impurity - MCE (%) BLQ BLQ BLQ BLQ 0.074 BLQ BLQ BLQ BLQ BLQImpurity - HP1 (%) 0.137 0.265 0.528 0.945 2.967 0.567 1.618 3.719 5.89214.427 Impurity - Dimer (%) BLQ BLQ BLQ 0.050 0.110 BLQ BLQ BLQ 0.0650.115 Single unknown (%) 0.112 0.105 0.086 0.054 0.112 0.057 BLQ ND NDND Total (%) 0.25 0.37 0.61 1.05 3.36 0.67 1.62 3.72 5.96 14.54Attribute Highest Concentration (0.6 mg/ml) Lowest Concentration (0.2mg/ml) Time Initial 1 hr 3 hrs 6 hrs 24 hrs Initial 1 hr 3 hrs 6 hrs 24hrs Formulation Treanda ™ 5 mg/ml admixed with 500 ml normal salineAssay (mg/ml) 0.566 0.558 0.544 0.527 0.454 0.193 0.191 0.185 0.1780.154 Assay (% Initial) 100.0 98.6 96.1 93.1 80.2 100.0 99.0 95.9 92.279.8 Impurity - MCE (%) 0.263 0.261 0.268 0.262 0.263 0.261 0.288 0.2770.250 0.276 Impurity - HP1 (%) 1.250 2.248 4.730 7.287 16.887 1.2312.241 4.770 7.462 17.504 Impurity - Dimer (%) 0.223 0.229 0.269 0.2790.326 0.188 0.185 0.178 0.176 0.252 Single unknown (%) 0.103 0.103 0.1010.097 0.081 0.077 0.079 0.103 0.083 0.066 Total (%) 1.97 2.97 5.50 8.0417.66 1.85 3.01 5.41 8.07 18.27 MCE—monochloroethyl derivative; HP1—monohydroxyl bendamustine

As shown in Table 9, Treanda™ when prepared as directed in the label(final concentration between 0.2-0.6 mg/ml) shows total degradation ofabout 5-6% in 3 hours at room temperature (corresponding to the roomtemperature stability claim in the label); monohydroxy bendamustine isthe main degradant. In contrast, the non-aqueous bendamustineformulations admixed in either 50 ml or 100 ml saline show totaldegradation of less than 5-6% over 6 hours at the lowest concentrationstested, indicating that these admixtures are significantly less prone todegradation. This stabilizing effect is particularly pronounced at thehigher concentrations (which are more typical), with chemical stabilityevident for 24 hours at these concentrations. The non-aqueousformulations of bendamustine thus offer better chemical stability thanTreanda™ when admixed into smaller volumes.

I claim:
 1. A method of treating cancer or malignant disease in asubject, comprising parenterally administering a volume of about 100 mlor less of a liquid composition comprising: a) from about 0.05 to about12.5 mg/ml of bendamustine or a pharmaceutically acceptable saltthereof; b) a solubilizer comprising polyethylene glycol and propyleneglycol, the polyethylene glycol being present in an amount of from about0.3 to about 45% by volume and the propylene glycol being present in anamount of from about 0.03 to about 5% by volume; c) a parenterallyacceptable diluent; and optionally d) an antioxidant; over a period ofless than or equal to about 10 minutes to the subject.
 2. The method ofclaim 1, wherein the subject is human.
 3. The method of claim 1, whereinthe concentration of the bendamustine or pharmaceutically acceptablesalt is from about 0.1 to about 6.0 mg/ml.
 4. The method of claim 1,wherein the concentration of the bendamustine or pharmaceuticallyacceptable salt is from about 0.05 to about 3.2 mg/ml.
 5. The method ofclaim 1, wherein the concentration of the bendamustine orpharmaceutically acceptable salt is from about 0.5 to about 5.6 mg/ml.6. The method of claim 1, wherein the amount of solubilizer is fromabout 0.5 to about 26.5% vol.
 7. The method of claim 1, wherein theamount of solubilizer is from about 0.2 to about 5% vol.
 8. The methodof claim 1, wherein the amount of solubilizer is from about 2.0 to about22.4% vol.
 9. The method of claim 1, where the polyethylene glycol isPEG
 400. 10. The method of claim 1, wherein the weight ratio ofpolyethylene glycol to propylene glycol is about 90:10.
 11. The methodof claim 5, wherein the weight ratio of polyethylene glycol to propyleneglycol is about 85:15.
 12. The method of claim 1, wherein the volumeadministered is about 50, 30 or 15 ml.
 13. The method of claim 1,wherein the antioxidant is monothioglycerol.
 14. The method of claim 10,wherein the volume administered is about 50 ml or less.
 15. The methodof claim 1, wherein the cancer or malignant disease is chroniclymphocytic leukemia.
 16. The method of claim 14, wherein thecomposition is administered intravenously in less than about 10 minuteson days 1 and 2 of a 28 day cycle.
 17. The method of claim 14, whereinthe composition is administered intravenously in less than about 10minutes on days 1 and 2 of a 28 day cycle.
 18. The method of claim 1,wherein the composition is administered for up to 6 cycles.
 19. Themethod of claim 14, wherein the volume of the composition administeredto the subject provides a bendamustine dosage amount ranging from about25 mg/m² to about 100 mg/m² to the subject.
 20. The method of claim 1,wherein the cancer or malignant disease is indolent B cell non-Hodgkin'slymphoma.
 21. The method of claim 15, wherein the composition isadministered intravenously in less than 10 minutes on days 1 and 2 of a21 day cycle.
 22. The method of claim 16, wherein the composition isadministered for up to 8 cycles.
 23. The method of claim 15, wherein thevolume of the composition administered to the subject provides abendamustine dosage amount ranging from about 60 mg/m² to about 120mg/m² to the subject.
 24. A method according to claim 1, wherein thebendamustine is present as the hydrochloride salt.